1-methyl-3-phenylpropylamine

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data of read across substances

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Principles of method if other than guideline:

WoE report is prepared based on short term toxicity to aquatic algae study.

GLP compliance:

not specified

Specific details on test material used for the study:

Name: 1-methyl-3-phenylpropylamine
Mol. formula: C10H15N
Molecular Weight: 149.236 g/mole
InChI: 1S/C10H15N/c1-9(11)7-8-10-5-3-2-4-6-10/h2-6,9H,7-8,11H2,1H3
Smiles: c1(CC[C@@H](C)N)ccccc1

Analytical monitoring:

yes

Details on sampling:

1. The test solution was prepared in aseptic condition. The test item was prepared by adding 100mg of test item in 200 ml of BBM to get the final concentration of 281.985mg/L. This stock solution was kept for sonication for 30 minutes to obtain a homogenous solution for the experiment. The test concentrations were chosen according to the available data of the test item. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 104cells/ml. Care was taken to have a homogeneous solution for the experiment.

Vehicle:

not specified

Details on test solutions:

1. The effect of the test item on the green algae Chlorella vulgaris culture was observed at nominal test concentration of 6.25mg/l, 12.5mg/l, 25mg/l, 50mg/l, 100 mg/l and 200 mg/l. All the six concentrations were in geometric series spaced by a factor of 2

2. The stock solution 100 g/l was prepared by dissolving colourless liquid in acetone. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water.

3. The stock solution 100 mg/L was prepared by dissolving transparent liquid in OECD growth medium. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture.

Test organisms (species):

other: Chlorella vulgaris and Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)

Details on test organisms:

1. TEST ORGANISM
- Common name: green alga
- Strain: Chlorella vulgaris
- Source (laboratory, culture collection): Laboratory
- Method of cultivation:Bold’s Basal Medium (BBM)

ACCLIMATION
- Culturing media and conditions (same as test or not):The medium to be used for the growth of algae was Bold’s Basal Medium (BBM). It is a medium composed of macronutrients, micronutrients, alkaline EDTA solution and Iron solution. Stock solution of each of these was prepared separately and then a complete medium was prepared and sterilized. De-ionized water was used to prepare the BBM
- Any deformed or abnormal cells observed: No

2, 3. TEST ORGANISM
- Strain: 86.81 SAG
- Source (laboratory, culture collection): Institute of botany of the ASCR
- Age of inoculum (at test initiation): 5x103 cells/ml

Test type:

static

Water media type:

freshwater

Total exposure duration:

72 h

Remarks on exposure duration:

±1 hour

Test temperature:

1. 24 °C ±2°C
2 and 3. 23±2°C

pH:

1. 6.6 to 7.7
2. Test at highest concentration: 7.8 (Change to 7.7 during tests)
3. The sample at concentration 80.0 mg/L : pH = 7.7 did not change during the test

Nominal and measured concentrations:

1. Six test concentration were: 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50mg/L, 100 mg/L and 200mg/L (Nominal)
2. 0, 0, 5, 10, 20, 40, 80 mg/l
3. 0, 5, 10, 20, 40 and 80 mg/L

Details on test conditions:

1. TEST SYSTEM
- Test vessel: Conical flasks
- Type (delete if not applicable): No data available
- Material, size, headspace, fill volume: Conical flasks of 100 ml size was used for the study.
- Aeration: No data available
- Type of flow-through (e.g. peristaltic or proportional diluter): No data available
- Renewal rate of test solution (frequency/flow rate): No data available
- Initial cells density: 54x105 cells/mL
- Control end cells density: No data available
- No. of organisms per vessel: 10000cells/ml
- No. of vessels per concentration (replicates): Two replicates for each test concentration
- No. of vessels per control (replicates): Three replicates for Control
- No. of vessels per vehicle control (replicates): No data available

GROWTH MEDIUM
- Standard medium used: No data available
- Detailed composition if non-standard medium was used: The medium to be used for the growth of algae was Bold’s Basal Medium (BBM). It is a medium composed of macronutrients, micronutrients, alkaline EDTA solution and Iron solution. Stock solution of each of these was prepared separately and then a complete medium was prepared and sterilized. De-ionized water was used to prepare the BBM.


OTHER TEST CONDITIONS
- Sterile test conditions: Yes
- Adjustment of pH: No data available
- Photoperiod: 16 Hour Light Period : 8 Hour Dark Period
- Light intensity and quality: continuous, uniform fluorescent illumination(1500Lux)
- Salinity (for marine algae): No data available

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: Spectrophotometer - The absorbance values of each test vessel and control vessel was noted at 680nm.The BBM was taken as blank for both control and test vessels. The absorbance value of each vessel was in line with the average specific growth rate.
- Chlorophyll measurement: No data available
- Other: Microscopic observations: The cultures were observed daily with the help of a microscope to verify a normal and healthy appearance of the algal culture and also to observe any abnormal appearance of the algae (as may be caused by the exposure of the test item). Apart from this, the cell count of each test vessel was also noted with the help of a microscope and haemocytometer.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: All the six concentrations were in geometric series spaced by a factor of 2.
- Justification for using less concentrations than requested by guideline: No data available
- Range finding study: No data available
- Test concentrations: Six test concentration were: 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50mg/L, 100 mg/L and 200mg/L (Nominal concentrations)
- Results used to determine the conditions for the definitive study: No data available


2. TEST SYSTEM
- Test vessel: 50 ml Glass vessel
- Type (delete if not applicable): closed (with air permeable stopper)
- Sample volume: 15 ml
- Initial cells density: 5x10(3) cells/ml
- No. of vessels per concentration (replicates): 3

GROWTH MEDIUM
- Standard medium used: yes

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: Continuous
- Light intensity and quality: 6000-8000 lx

3. TEST SYSTEM
- Test vessel: 50ml glass vessel
- Type : closed (with air-permeable stopper)
- Material, size, headspace, fill volume: 7.5 ml
- Initial cells density: 5x103 cells/mL
- No. of vessels per concentration (replicates): 3

GROWTH MEDIUM
- Standard medium used: Yes

OTHER TEST CONDITIONS
- Adjustment of pH: Without adjustment
- Photoperiod: Continuous
- Light intensity and quality: 6000 lx - 8000 lx

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (K2Cr2O7)

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 200 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

cell number

Remarks on result:

other: Other details not known

Duration:

72 h

Dose descriptor:

other: ErC50

Effect conc.:

71.1 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: 95 % CI was 62.6 - 80.8 mg/l

Duration:

72 h

Dose descriptor:

other: ErC50

Effect conc.:

90.8 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: 95 % CI was 82.6 - 99.7 mg/l

Results with reference substance (positive control):

2.Results with reference substance valid
- EC50: 0.75 mg/L

3. - Results with reference substance valid
- ErC50: 0.69 mg/L K2Cr2O7

Reported statistics and error estimates:

1.To obtain a quantitative concentration-response relationship by regression analysis, a line arizing transformation of the response data into probit was performed. Using the same, effective concentration (EC) was determined.
2. ErC50 is calculated from the inhibition of growth rate by non-linear regression using software Prism 4.0, GraphPad.

Validity criteria fulfilled:

yes

Conclusions:

1. After 72 hours of exposure to test item to various nominal test concentrations, EC50 was determine to be >200mg/l graphically and through probit analysis.
2. The median effective concentration (ErC50) for the test substance, in Desmodesmus subspicatus was determined to be 71.1 mg/L on the basis of effects on growth rate in a 72 hour study.
3. The median effective concentration (ErC50) for the test substance, in a freshwater algae Desmodesmus subspicatus was determined to be 90.8 mg/L on the basis of effects on growth rate in a 72 hour study.

Executive summary:

Short term toxicity of 1-methyl-3-phenylpropylamine (22374-89-6) on the growth and other biological activity of aquatic algae is predicted on the basis of it structurally and functionally similar read across chemicals. The studies are as mentioned below:

 

Short term toxicity study was performed to determine the toxic effect of the structurally and functionally similar read across chemicals on the algae. The effect of test item was studied on the growth of fresh water green alga Chlorella vulgaris. The study was conducted following OECD guideline 201- Alga growth inhibition test. The test concentration chosen for the study were 6.25mg/L, 12.5mg/L, 25mg/L, 50 mg/L, 100mg/L and 200mg/L. The test concentrations were prepared using stock solution of the test item using mineral media. The green alga was exposed to the test concentration for a period of 72 hours to observe average specific growth rate and % growth inhibition under the effect of the test item. EC50 calculated graphically through probit analysis was observed to be >200mg/L. Thus, based on this value, test chemical can be considered as non-toxic to aquatic organisms and thus cannot be classified as hazardous as per the CLP criteria.

 

Similar study was performed for another structurally and functionally similar read across chemicals. Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the substance. Test was conducted according to OECD Guideline 201. The stock solution 100 mg/l was prepared by dissolving colourless liquid in acetone. Test solutions of required concentration as were prepared by mixing the stock solution of the test sample with reconstituted test water. The test substance was tested at the concentrations 0, 0, 5, 10, 20, 40, 80 mg/L. Effects on growth rate were observed for 72 hours. The median effective concentration (ErC50) for the test substance, in Desmodesmus subspicatus was determined to be 71.1 mg/L for growth rate effects. This value indicates that the substance is likely to be hazardous to aquatic algae and can be classified as aquatic chronic 3 category as per the CLP criteria. But as the chemical was readily biodegradable thus it can be concluded that the chemical was nontoxic and not classified as per the CLP classification criteria.

 

Similarly third chemical was also supports the nontoxic nature of chemical which was structurally similar to the target chemical. Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the test substance according to OECD Guideline 201.The test substance was dissolved in OECD growth medium and tested at the concentrations 0, 5, 10, 20, 40 and 80 mg/L. Effects on the growth rate of the organism were studied. The median effective concentration (ErC50) for the test substance, in Desmodesmus subspicatus was determined to be 90.8 mg/L. But as the substance was found to be readily bio-degradable in water compartment. Thus considering only the value from CLP Criteria for aquatic classification of the substance, it is concluded that test substance does not exhibits toxicity to aquatic algae (Desmodesmus subspicatus) and not classified as per the CLP classification criteria.

 

Thus based on the overall studies for the structurally and functionally similar read across chemical for 1-methyl-3-phenylpropylamine (22374-89-6), it can be concluded that the test chemical was nontoxic and not classified as per the CLP classification criteria.  

Description of key information

Short term toxicity of 1-methyl-3-phenylpropylamine (22374-89-6) on the growth and other biological activity of aquatic algae is predicted on the basis of it structurally and functionally similar read across chemicals. The studies are as mentioned below:

 

Short term toxicity study was performed to determine the toxic effect of the structurally and functionally similar read across chemicals on the algae. The effect of test item was studied on the growth of fresh water green alga Chlorella vulgaris. The study was conducted following OECD guideline 201- Alga growth inhibition test. The test concentration chosen for the study were 6.25mg/L, 12.5mg/L, 25mg/L, 50 mg/L, 100mg/L and 200mg/L. The test concentrations were prepared using stock solution of the test item using mineral media. The green alga was exposed to the test concentration for a period of 72 hours to observe average specific growth rate and % growth inhibition under the effect of the test item. EC50 calculated graphically through probit analysis was observed to be >200mg/L. Thus, based on this value, test chemical can be considered as non-toxic to aquatic organisms and thus cannot be classified as hazardous as per the CLP criteria.

 

Similar study was performed for another structurally and functionally similar read across chemicals. Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the substance. Test was conducted according to OECD Guideline 201. The stock solution 100 mg/l was prepared by dissolving colourless liquid in acetone. Test solutions of required concentration as were prepared by mixing the stock solution of the test sample with reconstituted test water. The test substance was tested at the concentrations 0, 0, 5, 10, 20, 40, 80 mg/L. Effects on growth rate were observed for 72 hours. The median effective concentration (ErC50) for the test substance, in Desmodesmus subspicatus was determined to be 71.1 mg/L for growth rate effects. This value indicates that the substance is likely to be hazardous to aquatic algae and can be classified as aquatic chronic 3 category as per the CLP criteria. But as the chemical was readily biodegradable thus it can be concluded that the chemical was nontoxic and not classified as per the CLP classification criteria.

 

Similarly third chemical was also supports the nontoxic nature of chemical which was structurally similar to the target chemical. Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the test substance according to OECD Guideline 201.The test substance was dissolved in OECD growth medium and tested at the concentrations 0, 5, 10, 20, 40 and 80 mg/L. Effects on the growth rate of the organism were studied. The median effective concentration (ErC50) for the test substance, in Desmodesmus subspicatus was determined to be 90.8 mg/L. But as the substance was found to be readily bio-degradable in water compartment. Thus considering only the value from CLP Criteria for aquatic classification of the substance, it is concluded that test substance does not exhibits toxicity to aquatic algae (Desmodesmus subspicatus) and can be classified as aquatic chronic 3 as per the CLP classification criteria.

 

Thus based on the overall studies for the structurally and functionally similar read across chemical for 1-methyl-3-phenylpropylamine (22374-89-6), it can be concluded that the test chemical was nontoxic and not classified as per the CLP classification criteria.  

Key value for chemical safety assessment

EC50 for freshwater algae:

200 mg/L

Additional information

Summarized result for the toxicity of chemical, on the growth of aquatic algae and cyanobacteria. Short term toxicity of 1-methyl-3-phenylpropylamine (22374-89-6) on the growth and other biological activity of aquatic algae is predicted on the basis of it experimentally structurally and functionally similar read across chemicals. The studies are as mentioned below:

 

Short term toxicity study was performed to determine the toxic effect of the structurally and functionally similar read across chemicals on the algae. The effect of test item was studied on the growth of fresh water green alga Chlorella vulgaris. The study was conducted following OECD guideline 201- Alga growth inhibition test. The test concentration chosen for the study were 6.25mg/L, 12.5mg/L, 25mg/L, 50 mg/L, 100mg/L and 200mg/L. The test concentrations were prepared using stock solution of the test item using mineral media. The green alga was exposed to the test concentration for a period of 72 hours to observe average specific growth rate and % growth inhibition under the effect of the test item. EC50 calculated graphically through probit analysis was observed to be >200mg/L. Thus, based on this value, test chemical can be considered as non-toxic to aquatic organisms and thus cannot be classified as hazardous as per the CLP criteria.

 

Similar study was performed for another structurally and functionally similar read across chemicals. Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the substance. Test was conducted according to OECD Guideline 201. The stock solution 100 mg/l was prepared by dissolving colourless liquid in acetone. Test solutions of required concentration as were prepared by mixing the stock solution of the test sample with reconstituted test water. The test substance was tested at the concentrations 0, 0, 5, 10, 20, 40, 80 mg/L. Effects on growth rate were observed for 72 hours. The median effective concentration (ErC50) for the test substance, in Desmodesmus subspicatus was determined to be 71.1 mg/L for growth rate effects. This value indicates that the substance is likely to be hazardous to aquatic algae and can be classified as aquatic chronic 3 category as per the CLP criteria. But as the chemical was readily biodegradable thus it can be concluded that the chemical was nontoxic and not classified as per the CLP classification criteria.

 

Similarly third chemical was also supports the nontoxic nature of chemical which was structurally similar to the target chemical. Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the test substance according to OECD Guideline 201.The test substance was dissolved in OECD growth medium and tested at the concentrations 0, 5, 10, 20, 40 and 80 mg/L. Effects on the growth rate of the organism were studied. The median effective concentration (ErC50) for the test substance, in Desmodesmus subspicatus was determined to be 90.8 mg/L. Thus considering only the value from CLP Criteria for aquatic classification of the substance, it is concluded that test substance does not exhibits toxicity to aquatic algae (Desmodesmus subspicatus) and can be classified as aquatic chronic 3 as per the CLP classification criteria.

 

Thus based on the overall studies for the structurally and functionally similar read across chemical for 1-methyl-3-phenylpropylamine (22374-89-6), it can be concluded that the test chemical was nontoxic and not classified as per the CLP classification criteria.